Rapid rates of sperm DNA damage after activation in tench (Tinca tinca: Teleostei, Cyprinidae) measured using a sperm chromatin dispersion test

Lopez-Fernandez, C, Gage, MJG, Arroyo, F, Gosalbez, A, Larran, AM, Fernandez, JL and Gosalvez, J (2009) Rapid rates of sperm DNA damage after activation in tench (Tinca tinca: Teleostei, Cyprinidae) measured using a sperm chromatin dispersion test. Reproduction, 138 (2). pp. 257-266. ISSN 1470-1626

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Abstract

Spermatozoal haplotypic DNA is prone to damage, leading to male fertility problems. So far, the assessment of sperm DNA breakage has been challenging because protamines render the nuclear chromatin highly compacted. Here, we report the application of a new test to quantify DNA fragmentation in spermatozoa of an externally fertilizing teleost fish. The sperm chromatin dispersion (SCD) test uses a species-specific lysing solution to generate controlled protein depletion that, followed by DNA-specific fluorescent labelling, allows an easy morphological discrimination between nuclei affected by DNA damage. Using tench (Tinca tinca) as our model, we first trialled the test against established, but more technically demanding, assays employing in situ nick translation (ISNT) and the comet assay. The SCD test showed high concordance with ISNT, comet assay measures and a chromatin-swelling test, confirming the application of this straightforward SCD technique to various aspects of reproductive biology. Second, we examined between-male variation in DNA damage, and measured changes through time following spermatozoal activation. Between-male variation in the basal levels of average DNA damage ranged from 0 to 20% of sperm showing damage, and all showed increases in DNA fragmentation through time (0–60 min). The rates of DNA damage increase are the fastest so far recorded in sperm for a living organism, and may relate to the external fertilization mode. Our findings have relevance for broodstock selection and optimizing IVF protocols routinely used in modern aquaculture.

Item Type: Article
Faculty \ School: Faculty of Science > School of Biological Sciences
University of East Anglia > Faculty of Science > Research Centres > Centre for Ecology, Evolution and Conservation
University of East Anglia > Faculty of Science > Research Groups > Organisms and the Environment
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Depositing User: Deborah Clemitshaw
Date Deposited: 18 May 2011 11:08
Last Modified: 25 Jul 2018 04:55
URI: https://ueaeprints.uea.ac.uk/id/eprint/30769
DOI: 10.1530/REP-09-0105

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